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: A case-control design study was used to investigate the association of maternal HIV-1 phenotype in MT-2 cells at or near the time of delivery with perinatal transmission of HIV-1, controlling for maternal CD4 percentage and duration of rupture of membranes, in 48 transmitting and 96 nontransmitting HIV-1-infected mothers who gave birth between 1990 and 1995. The nonsyncytium-indueing (NSI) phenotype was more commonly seen in transmitting mothers compared with nontransmitting mothers (90% vs. 75%, p = .04). In a multivariable logistic regression model, the following maternal characteristics were significantly associated with HIV transmission: NSI phenotype (odds ratio [OR] = 6.08; 95% confidence interval [CI]: 1.73- 21.35) and log10 viral load (OR = 2.11; CI: 1.19-3.74). Finally, the association of NSI phenotype with transmission was stronger in transmitting women who received azidothymidine during pregnancy compared with transmitters who did not.